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PMID: 10339558 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Regulation of the OxyR transcription factor by hydrogen peroxide and the cellular thiol-disulfide status.

Aslund F, Zheng M, Beckwith J, Storz G

Abstract

The Escherichia coli transcription factor OxyR is activated by the formation of an intramolecular disulfide bond and subsequently is deactivated by enzymatic reduction of the disulfide bond. Here we show that OxyR can be activated by two possible pathways. In mutants defective in the cellular disulfide-reducing systems, OxyR is constitutively activated by a change in the thiol-disulfide redox status in the absence of added oxidants. In wild-type cells, OxyR is activated by hydrogen peroxide. By monitoring the presence of the OxyR disulfide bond after exposure to hydrogen peroxide in vivo and in vitro, we also show that the kinetics of OxyR oxidation by low concentrations of hydrogen peroxide is significantly faster than the kinetics of OxyR reduction, allowing for transient activation in an overall reducing environment. We propose that the activity of OxyR in vivo is determined by the balance between hydrogen peroxide levels and the cellular redox environment.

MeSH Terms
Alkaline Phosphatase/metabolism Bacterial Proteins/metabolism DNA-Binding Proteins Escherichia coli/genetics,metabolism Escherichia coli Proteins Genotype Glutathione/metabolism Glutathione Disulfide/metabolism Hydrogen Peroxide/pharmacology Kinetics Models, Chemical Oxidation-Reduction Repressor Proteins/metabolism Transcription Factors/metabolism
Chemicals
Bacterial Proteins DNA-Binding Proteins Escherichia coli Proteins Repressor Proteins Transcription Factors oxyR protein, E coli Hydrogen Peroxide Alkaline Phosphatase Glutathione Glutathione Disulfide
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Aslund F
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
Zheng M
Beckwith J
Storz G
References (22)
22 references, click to expand
  1. The in vivo distribution of oxidized and reduced thioredoxin in Escherichia coli.
    J Biol Chem. 1982 Jun 25;257(12):6926-30 PMID: 7045097
  2. Molecular and cellular aspects of thiol-disulfide exchange.
    Adv Enzymol Relat Areas Mol Biol. 1990;63:69-172 PMID: 2407068
  3. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
    Anal Biochem. 1987 Apr;162(1):156-9 PMID: 2440339
  4. The non-flavin redox center of the streptococcal NADH peroxidase. II. Evidence for a stabilized cysteine-sulfenic acid.
    J Biol Chem. 1989 Jul 25;264(21):12330-8 PMID: 2501303
  5. Transcriptional regulator of oxidative stress-inducible genes: direct activation by oxidation.
    Science. 1990 Apr 13;248(4952):189-94 PMID: 2183352
  6. Oxidized redox state of glutathione in the endoplasmic reticulum.
    Science. 1992 Sep 11;257(5076):1496-502 PMID: 1523409
  7. Mutations that allow disulfide bond formation in the cytoplasm of Escherichia coli.
    Science. 1993 Dec 10;262(5140):1744-7 PMID: 8259521
  8. Effects of peroxides on susceptibilities of Escherichia coli and Mycobacterium smegmatis to isoniazid.
    Antimicrob Agents Chemother. 1994 Aug;38(8):1829-33 PMID: 7986015
  9. OxyR regulon.
    Methods Enzymol. 1994;234:217-23 PMID: 7528872
  10. Analysis of the kinetic mechanism of enterococcal NADH peroxidase reveals catalytic roles for NADH complexes with both oxidized and two-electron-reduced enzyme forms.
    Biochemistry. 1995 Oct 31;34(43):14114-24 PMID: 7578008
  11. Distribution of thiols in microorganisms: mycothiol is a major thiol in most actinomycetes.
    J Bacteriol. 1996 Apr;178(7):1990-5 PMID: 8606174
  12. The levels of ribonucleotide reductase, thioredoxin, glutaredoxin 1, and GSH are balanced in Escherichia coli K12.
    J Biol Chem. 1996 Aug 9;271(32):19099-103 PMID: 8702583
  13. Homeostatic regulation of intracellular hydrogen peroxide concentration in aerobically growing Escherichia coli.
    J Bacteriol. 1997 Jan;179(2):382-8 PMID: 8990289
  14. The role of the thioredoxin and glutaredoxin pathways in reducing protein disulfide bonds in the Escherichia coli cytoplasm.
    J Biol Chem. 1997 Jun 20;272(25):15661-7 PMID: 9188456
  15. A small, stable RNA induced by oxidative stress: role as a pleiotropic regulator and antimutator.
    Cell. 1997 Jul 11;90(1):43-53 PMID: 9230301
  16. Activation of the OxyR transcription factor by reversible disulfide bond formation.
    Science. 1998 Mar 13;279(5357):1718-21 PMID: 9497290
  17. Different mechanisms of thioredoxin in its reduced and oxidized forms in defense against hydrogen peroxide in Escherichia coli.
    Free Radic Biol Med. 1998 Mar 1;24(4):556-62 PMID: 9559867
  18. In vivo transcription of nrdAB operon and of grxA and fpg genes is triggered in Escherichia coli lacking both thioredoxin and glutaredoxin 1 or thioredoxin and glutathione, respectively.
    J Biol Chem. 1998 Jul 17;273(29):18382-8 PMID: 9660805
  19. sigmaR, an RNA polymerase sigma factor that modulates expression of the thioredoxin system in response to oxidative stress in Streptomyces coelicolor A3(2).
    EMBO J. 1998 Oct 1;17(19):5776-82 PMID: 9755177
  20. Bacterial senescence: stasis results in increased and differential oxidation of cytoplasmic proteins leading to developmental induction of the heat shock regulon.
    Genes Dev. 1998 Nov 1;12(21):3431-41 PMID: 9808629
  21. Chaperone activity with a redox switch.
    Cell. 1999 Feb 5;96(3):341-52 PMID: 10025400
  22. Determination of glutathione and glutathione disulfide in biological samples.
    Methods Enzymol. 1985;113:548-55 PMID: 4088074
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1999-05-25
Pages
6161-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC26852
Subset
IM
Grants
NIGMS NIH HHS · R01 GM041883 · United States
NIGMS NIH HHS · R01 GM055090 · United States
NIGMS NIH HHS · GM41883 · United States
NIGMS NIH HHS · GM55090 · United States
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